Subcellular Protein Fractionation Kit for Cultured Cells
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Subcellular Protein Fractionation Kit for Cultured Cells
Thermo Scientific™

Subcellular Protein Fractionation Kit for Cultured Cells

Thermo Scientific Subcellular Protein Fractionation Kit for Cultured Cells enables segregation and enrichment of proteins from five different cellular compartments.FeaturesRead more
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Catalog number 78840
Price (USD)
598.00
Each
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Price (USD)
598.00
Each
Add to cart
Thermo Scientific Subcellular Protein Fractionation Kit for Cultured Cells enables segregation and enrichment of proteins from five different cellular compartments.

Features of Subcellular Protein Fractionation Kit for Cultured Cells:

Concise—extract functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions from mammalian cultured cells in less than 3 hours
Convenient—perform a simple procedure without using ultracentrifugation over gradients
Complete—obtain cytoplasmic, membrane, soluble nuclear, chromatin-bound and cytoskeletal protein fractions from a single kit
Cultured Cells—developed and optimized for use with cultured mammalian cells; for tissue samples use the Subcellular Fractionation Kit for Tissues
Compatible—use extracts for downstream applications such as protein assays, Western blotting, protein assays, electrophoretic mobility shift assays, reporter-gene and enzyme-activity assays.

Applications for subcellular fractionation:
• Extract and enrich proteins from different cellular compartments
• Isolate cytoplasmic, nuclear, membrane and cytoskeletal proteins from the same cells
• Profile cells for proteins from the cytoplasm, nucleus, membrane and cytoskeleton
• Study protein translocation

Fractionation of subcellular proteins enables protein localization assessment and protein enrichment from specific cellular compartments. Thermo Scientific Subcellular Protein Fractionation Kit includes a combination of reagents for stepwise lysis of cells into functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than 3 hours. Extracts from each subcellular compartment generally have less than 15% contamination between fractions, which is sufficient purity for most experiments studying protein localization and redistribution.

The Subcellular Protein Fractionation Kit for Cultured Cells contains four extraction buffers, a stabilized nuclease and Thermo Scientific Halt Protease Inhibitor Cocktail. Each kit has enough reagents to fractionate 50 cell pellets of 20 μL each, equivalent to approximately 2g of cell paste. (For tissue samples, use our Subcellular Protein Fractionation Kit for Tissue.)

The first kit reagent, when added to a cell pellet, causes selective membrane permeabilization, releasing soluble cytoplasmic contents. The second reagent dissolves plasma, mitochondria and ER-golgi membranes but does not solubilize the nuclear membranes. After recovering intact nuclei by centrifugation, a third reagent yields the soluble nuclear extract. An additional nuclear extraction with micrococcal nuclease is performed to release chromatin-bound nuclear proteins. The recovered insoluble pellet is then extracted with the final reagent to isolate cytoskeletal proteins.

Related Products
Subcellular Protein Fractionation Kit for Tissues
Benchtop Centrifuges
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeSubcellular Protein Fractionation Kit
FormLiquid
BufferLysis Buffer
Reagent TypeCell Lysis Buffer, Detergent Solution, Subcellular Fractionation
Sufficient For50 Samples
Quantity1 Kit
Unit SizeEach
Contents & Storage
Upon receipt store kit at -20°C, or store individual components as indicated above. Kit is shipped with dry ice.

Frequently asked questions (FAQs)

What products are available for the isolation of subcellular components?

You can choose from several types of Dynabeads magnetic beads for coating with antibody against the subcellular component of choice. We currently do not offer any ready-to-use Dynabeads magnetic beads against a particular subcellular component, and recommend that you choose the beads that work best with your antibody and your application.

The following groups of Dynabeads magnetic beads can be used for this purpose:

(1) Surface Activated Dynabeads magnetic beads
Antibodies from any origin can be conjugated directly to these uncoated beads through covalent bonds. For better orientation of primary antibody on the beads, a secondary antibody (linker antibody) can be coated on the beads first. The Surface Activated Dynabeads magnetic beads are provided in different sizes (4.5, 2.8 and 1 micron) and the larger ones are mostly preferred for this application.

(2) Secondary Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are pre-coated with a secondary antibody and depending on their specificity, they can be easily coated with your primary antibody within a short time period. These beads are provided in two sizes, 4.5 and 2.8 micron. In addition to these beads, our Dynabeads magnetic beads Protein A or Protein G can also be easily coated with antibody from several species. Please see the following links for information about all available Secondary coated Dynabeads magnetic beads and Dynabeads magnetic beads Protein A and Protein G.

(3) Streptavidin Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are coated with streptavidin and require a biotinylated antibody. These beads are provided in two different sizes, 2.8 and 1 micron.

Find more information about Dynabeads magnetic beads at http://www.thermofisher.com/Dynabeads magnetic beads.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can you provide the shelf-life for the Subcellular Protein Fractionation Kit for Cultured Cells?

The Subcellular Protein Fractionation Kit for Cultured Cells is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can I use the Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for subsequent co-immunoprecipitation (co-IP) analysis of different nuclear proteins?

Yes, the nuclear fraction is compatible with co-immunoprecipitation (co-IP) after dialysis, desalting, or 50:50 dilution. The cytoplasmic and membrane fractions are compatible with co-IP directly after extraction and the pellet fraction is compatible with co-IP after detergent removal.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Will M-PER Mammalian Protein Extraction Reagent extract membrane or cytoskeletal proteins?

M-PER Mammalian Protein Extraction Reagent can extract some membrane or cytoskeletal proteins, but the extraction efficiency is not consistent. The reagent was not intended to specifically extract these proteins. We recommend using Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) for membrane protein extraction or Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for compartmental extraction.

Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.

What kits do you offer for cell fractionation?

We offer the following kits for cell fractionation:

Mem-PER Plus Membrane Protein Extraction Kit, Cat. No. 89842
NE-PER Nuclear and Cytoplasmic Extraction Reagents, Cat. No. 78835
Subcellular Protein Fractionation Kit for Cultured Cells, Cat. No. 78840
Subcellular Protein Fractionation Kit for Tissues, Cat. No. 87790
Mitochondrial Isolation Kit for Cultured Cells, Cat. No. 89874
Mitochondrial Isolation kit for Tissue, Cat. No. 89801
Lysosome Enrichment Kit for Tissues and Cultured Cells, Cat. No. 89839

Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.